Antioxidant vitamins and lysophospholipids are critical for inducing mouse spermatogenesis under organ culture conditions

Sanjo, Hiroyuki; Yao, Tatsuma; Katagiri, Kumiko; Sato, Takuya; Matsumura, Takafumi; Komeya, Mitsuru; Yamanaka, Hidetoshi; Yao, Masahiro; Matsuhisa, Akio; Asayama, Yuta; Ikeda, Kazutaka; Kano, Kuniyuki; Aoki, Junken; Arita, Makoto; Ogawa, Takehiko

doi:10.1096/fj.202000245r

Cited by 17 publications

(20 citation statements)

References 55 publications

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“…These substances were effective in mouse IVS, too. We found in our previous studies that hormones, antioxidants, and lysophospholipids examined in this study had a definite effect on improving IVS in mice 21 , 22 . Thus, their effects are not specific to rats, but are common to mice as well.…”

Section: Discussionsupporting

confidence: 56%

“…In our previous study, lysophosoplipids was shown to be effective in inducing/promoting mouse IVS 22 . So, we investigated if the lysophopholipids are effective to rat IVS as well.…”

Section: Resultsmentioning

confidence: 89%

“…We also added hormones, consisting of testosterone (T), triiodothyronine (T 3 ), luteinizing hormone (LH), and follicle stimulating hormone (FSH), which was named 4 hormones (4Hs). These supplements had been shown to be effective in promoting mouse spermatogenesis 21 , 22 . Secondly, we applied silicone cover ceiling method, which used polydimethylsiloxane (PDMS) ceiling (PC) chip having 160 µm-depth square dent, to hold the testis tissue in the dent space making it thin and flat on the agarose gel (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Then, we found that antioxidative substances, α-tocopherol, ascorbic acid, and glutathione, and lysophospholipids, particularly lysophosphatidylcholine (LPC), lysophosphatidic acid (LPA), and lysophosphatidylserine (LysoPS), were critically effective for the induction and promotion of mouse spermatogenesis. By supplementing these substances in the culture medium, mouse spermatogenesis was induced in neonatal mouse testis even without using AlbuMAX 21 , 22 .…”

Section: Introductionmentioning

confidence: 99%

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Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control

Matsumura

Sato

Abe

et al. 2021

Sci Rep

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In vitro spermatogenesis (IVS) using air–liquid interphase organ culture method is possible with mouse testis tissues. The same method, however, has been hardly applicable to animals other than mice, only producing no or limited progression of spermatogenesis. In the present study, we challenged IVS of rats with modifications of culture medium, by supplementing chemical substances, including hormones, antioxidants, and lysophospholipids. In addition, reducing oxygen tension by placing tissues in an incubator of lower oxygen concentration and/or applying silicone cover ceiling on top of the tissue were effective for improving the spermatogenic efficiency. Through these modifications of the culture condition, rat spermatogenesis up to round spermatids was maintained over 70 days in the cultured tissue. Present results demonstrated a significant progress in rat IVS, revealing conditions commonly favorable for mice and rats as well as finding rat-specific optimizations. This is an important step towards successful IVS in many animal species, including humans.

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Section: Discussionsupporting

confidence: 56%

“…In our previous study, lysophosoplipids was shown to be effective in inducing/promoting mouse IVS 22 . So, we investigated if the lysophopholipids are effective to rat IVS as well.…”

Section: Resultsmentioning

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control

Matsumura

Sato

Abe

et al. 2021

Sci Rep

Self Cite

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“…Lipids are critical components in spermatogenesis. For example, albumin-bound lipids such as lysophospholipids are critical for inducing mouse spermatogenesis even under organ culture conditions 17 . Phospholipids containing PUFAs, especially DHA, are important for the formation and function of sperm 18 – 20 .…”

Section: Discussionmentioning

confidence: 99%

Abnormal male reproduction and embryonic development induced by downregulation of a phospholipid fatty acid-introducing enzyme Lpgat1 in zebrafish

Shibata

Kawana

Nishino

et al. 2022

Sci Rep

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Phospholipids in the membrane consist of diverse pairs of fatty acids bound to a glycerol backbone. The biological significance of the diversity, however, remains mostly unclear. Part of this diversity is due to lysophospholipid acyltransferases (LPLATs), which introduce a fatty acid into lysophospholipids. The human genome has 14 LPLATs and most of them are highly conserved in vertebrates. Here, we analyzed the function of one of these enzymes, lysophosphatidylglycerol acyltransferase 1 (Lpgat1), in zebrafish. We found that the reproduction of heterozygous (lpgat1+/−) male mutants was abnormal. Crosses between heterozygous males and wild-type females produced many eggs with no obvious cleavage, whereas eggs produced by crosses between heterozygous females and wild-type males cleaved normally. Consistent with this, spermatozoa from heterozygous males had reduced motility and abnormal morphology. We also found that the occurrence of lpgat1 homozygous (lpgat1−/−) mutants was far lower than expected. In addition, downregulation of lpgat1 by morpholino antisense oligonucleotides resulted in severe developmental defects. Lipidomic analysis revealed that selective phospholipid species with stearic acid and docosahexaenoic acid were reduced in homozygous larvae and spermatozoa from heterozygotes. These results suggest that the specific phospholipid molecular species produced by Lpgat1 have an essential role in sperm fertilization and in embryonic development.

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Role of autophagy in lysophosphatidylcholine‐induced apoptosis in mouse Leydig cells

Zeng

Yang

et al. 2022

Environmental Toxicology

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Lysophosphatidylcholine (LPC), a major class of glycerophospholipids ubiquitously present in most tissues, plays a dominant role in many diseases, while it is still unknown about the potential mechanism of LPC affecting the testicular Leydig cells.In the present study, mouse TM3 Leydig cells in vitro were treated with LPC for 48 h. LPC was found to significantly induce apoptosis and oxidative stress of mouse TM3 Leydig cells; while inhibition of oxidative stress by N-acetyl-L-cysteine, an inhibitor of oxidative stress, could rescue the induction of apoptosis, indicating that LPC induced apoptosis of mouse TM3 Leydig cells via oxidative stress. Interestingly, LPC was showed to inhibit autophagy; however, induction of autophagy by rapamycin significantly alleviated the induction of apoptosis by LPC. Taken together, oxidative stress was involved in LPC-induced apoptosis of mouse TM3 Leydig cells, and autophagy might play a protective role in LPC-induced apoptosis.

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Antioxidant vitamins and lysophospholipids are critical for inducing mouse spermatogenesis under organ culture conditions

Abstract: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

Cited by 17 publications

References 55 publications

Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control

Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control

Abnormal male reproduction and embryonic development induced by downregulation of a phospholipid fatty acid-introducing enzyme Lpgat1 in zebrafish

Role of autophagy in lysophosphatidylcholine‐induced apoptosis in mouse Leydig cells

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